微弧氧化处理对钛表面MC3T3-E1细胞生物学行为的影响  

Effects of micro-arc oxidation on the biological behaviors of MC3T3-E1 cells on titanium

作　　者：李玉梅[1] 査年保 马威[2] 赵铱民[3] 

机构地区：[1]湖北文理学院附属医院.襄阳市中心医院口腔科,湖北襄阳441021 [2]第四军医大学口腔医学院种植科,陕西西安710032 [3]第四军医大学口腔医学院修复科,陕西西安710032

出　　处：《上海口腔医学》2015年第5期551-556,共6页Shanghai Journal of Stomatology

摘　　要：目的 :使用微弧氧化(micro-arc oxidation,MAO)方法对纯钛圆片进行表面处理,观察其对MC3T3-E1成骨前体细胞生物学行为的影响。方法:使用分级电压增强过程对纯钛试件进行MAO处理,场发射扫描电镜观察试样表面形态,测量不同表面的接触角。MC3T3-E1细胞接种于试样表面并进行体外培养,MTT法检测60、120 min不同表面的细胞黏附量;培养4 h后,激光共聚焦扫描显微镜观察附着细胞的骨架蛋白;24、72、120和168 h后,检测细胞增殖情况;培养第16天,实时荧光定量PCR(q PCR)检测细胞成骨相关基因的表达。采用SPSS16.0软件包对数据进行统计学分析。结果:MAO处理后,可在纯钛表面形成一层多孔氧化层,MAO钛表面水和甘油的接触角显著小于抛光钛表面。在2个时间点,细胞在MAO钛表面的黏附均高于抛光钛表面。肌动蛋白染色提示,在MAO钛表面,细胞伸展良好。从培养第72 h开始,细胞在MAO钛表面的增殖略高于抛光钛表面,但差异无显著性。q PCR结果表明,RUNX2和ALP基因的m RNA水平在MAO钛表面和抛光钛表面之间并无显著差异。结论:MAO处理可促进MC3T3-E1细胞在纯钛表面的早期黏附,但RUNX2和ALP基因的m RNA表达水平在各组间无显著差异。PURPOSE： This study was intended to modify the surface of pure titanium by micro-arc oxidation （MAO）, and to investigate the effects of MAO process on the biological behavior of MC3T3-E1 osteoblastic cells. METHODS： MAO treatment of specimens were carried out using a staggered voltage boost procedure. The surface topography of the prepared specimens were observed by field emission scanning electron microscopy. Contact angle measurements were tested on a contact angle measuring system. MC3T3-E1 cells were cultured on specimens and the number of adhesion cells at 60 and 120 min were investigated by MTT. After 4 h of culture, cytoskeleton of the attached cells were examined using laser confocal scanning microscope. After 24, 72, 120 and 168 h of post seeding, cell proliferation were assessed using MTT assay. On day 16 of culture, the expressions of osteogenesis-related genes were analyzed through real time fluorescence quantitative polymerase chain reaction （qPCR）. The data was analyzed using SPSS 16.0 software package. RESULTS： A porous oxide layer was grown on pure titanium suhstrates via MAO process. The contact angles for water and glycerol on the MAO surface were smaller than polished surface. At 2 culture times, the MAO surface showed significantly higher cell adhesion than polished surface. Actin staining indicated that the cells spread well on the MAO surface. At 72, 120 and 168 h, better cell proliferation were seen for MAO surfaces compared with that on the polished surface, but there was no significant difference. The qPCR showed that no obvious variations in gene expression of RUNX2 and ALP by MC3T3-E1 cells were observed on 2 different surfaces. CONCLUSIONS： Compared with polished surface, better adhesion of MC3T3-E1 cells are observed on MAO surface. However, no obvious change in gene expression of RUNX2 and ALP were observed between the MAO surface and polished surface.

关 键 词：钛 微弧氧化 细胞黏附 基因表达 

分 类 号：R783.1[医药卫生—口腔医学]